Large scale synthesis of dinucleoside monophosphates catalyzed by ribonuclease from Aspergillus clavatus

Abstract

A gram scale enzymatic synthesis of eight dinucleoside monophosphates (ApC, ApU, CpC, CpU, GpC, GpU, UpC, and UpU) is described. The synthesis involves a reaction between the appropriate ribonucleoside-2',3'-cyclic phosphates and cytidine or uridine in the presence of ribonuclease from Aspergillus clavatus at 30°C. The enzyme is removed from the reaction mixture by chromatography on Bio-Gel P4, and the dinucleoside monophosphate is further purified by chromatography on a DEAF,-Sephadex A-25, column.A procedure for the large scale preparation of the ribonuclease from Aspergillus clavatus is also described.

Phospholipase‐C from Bacillus cereus: Production, purification, and properties

“…The diglycerides were extracted from the plates with ether:ethanol, 3: 1, and ester bonds were determined by the hydroxamate method as described by Snyder et al 15 Ribonuclease activity was determined according to a previously published procedure. 16 Proteolytic activity was determined as described by L a s k o~s k i , '~ and inorganic phosphate by the method of Fiske and SubbaRow.ls Acid production was determined with a Model TTT a1 Titrator (Radiometer, Copenhagen).…”